effect of different carbon sources on in vitro shoot regeneration of petunia hybrid hort. (vilm)

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Effect of Different Carbon Sources on Petunia Hybrid

Assia Zahoor1Student, 2Ph.D Research Scholar

1,2,3Plant Tissue Culture Research Laboratory, Department of Botany,University of Kashmir, Hazratbal, Srinagar, India

ABSTRACT: The present study was undertaken in order to develop an efficient in vitro propagation protocol for hybrid using leaf as an explant. The effects of different concentrations of five different carbon sources i.e., sucrose, glucose, mannose, arabinosemaltose was investigated on shoot regeneration of Petunia hybrid. Shoot regeneration occurred on MS medium supplemented with sucrose, glucose and maltose. No growth was obtained on MS medium when mannose and arabinose were used as carbon source. Best shoot regeneration with maximum shoot length and maximum number of shoots was obtained on MS medium supplemented with 3% carbon within 11 days with 70% culture response. Shoot regeneration was also obtained on MS medium supplemented with glucose and maltose within 18 and 24 days with 50 and 30% culture response respectively. Keywords: Explant, Maltose, MS medium, Petunia hybrid, Regeneration 1. INTRODUCTION

Petunia is the herbaceous ornamental plant belonging to the family Solanaceae and consists of 35 species. The geographical distribution includes temperate and sub tropical regions of Argentina, Uruguay, Paraguay, Bolivia and Brazil (Ando et al., 2005). The basic Petunia flower is funnel shaped. Leaves and stems are sticky to the touch and have a distinctive odor.high economic importance in global horticulture and is grown for decorative purposes in gardens and landscape design projects, as house plants, for cut flowers and specimen display. Petunias

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of Different Carbon Sources on in Vitro Shoot Regeneration oPetunia Hybrid Hort. (Vilm)

Assia Zahoor1, Jasfeeda Qadir2, Dr. Seema Singh3 Ph.D Research Scholar, 3Sr. Assistant Professor

Plant Tissue Culture Research Laboratory, Department of Botany,University of Kashmir, Hazratbal, Srinagar, India

The present study was undertaken in order to develop propagation protocol for Petunia

using leaf as an explant. The effects of different concentrations of five different carbon sources i.e., sucrose, glucose, mannose, arabinose and maltose was investigated on shoot regeneration of

. Shoot regeneration occurred on MS medium supplemented with sucrose, glucose and maltose. No growth was obtained on MS medium when mannose and arabinose were used as carbon

shoot regeneration with maximum shoot length and maximum number of shoots was obtained on MS medium supplemented with 3% carbon within 11 days with 70% culture response. Shoot regeneration was also obtained on MS medium

e within 18 and 24 days with 50 and 30% culture response

Explant, Maltose, MS medium, Petunia

is the herbaceous ornamental plant belonging and consists of 35 species.

The geographical distribution includes temperate and sub tropical regions of Argentina, Uruguay, Paraguay,

., 2005). The basic flower is funnel shaped. Leaves and stems are

uch and have a distinctive odor. It has high economic importance in global horticulture and is grown for decorative purposes in gardens and landscape design projects, as house plants, for cut

Petunias are a great

flowering bedding plant to add to landscape prior to the end of winter/beginning of spring. Petunia hybridais a petunia plant “nothospecies’’ (hybrid), which encompasses all hybrid species of petunia between P.axillarisand P.integrifolia. Petunia hybrid were originally produced by hybridization between P.axillaris (the large white or night and P.integrifolia (the violet-flowered petunia) (Skin, 1984). Petunia is used as a model plant due to its early growth habit and relativelyasexual propagation from cuttings, callus or protoplasts, availability of large sets of mutants, amenity for cytogenetic analysis. Besides ornamental value, Petunia also possesses medicinal value like it has antimicrobial activity, antioxidant activity and insecticidal activity. MATERIALS AND METHODS Washing and sterilization of explantsFor standardization of in vitro of Petunia hybrid were collected from plants growing in Kashmir University Botanical Garden. In order to remove dirt and dust, the explants were thoroughly washed under running tap water followed by washing with detergent labolene and surfactant tweendetergent was removed by washing the explants with double distilled water. Then the leaf explants were treated with chemical sterilants2% sodium hypochlorite for 5 minutes under laminar air flow hood. This was followed by washing with autoclaved double distilled water. Finally the leaf explants were inoculated on Murashige and Skoog (1962) medium.

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Scientific (IJTSRD)

International Open Access Journal

Shoot Regeneration of

Plant Tissue Culture Research Laboratory, Department of Botany,

to add to landscape prior to the end of winter/beginning of spring. Petunia hybrida is a petunia plant “nothospecies’’ (hybrid), which encompasses all hybrid species of petunia between P.axillarisand P.integrifolia. Petunia hybrid plants were originally produced by hybridization between

(the large white or night – scented petunia) flowered petunia) (Skin,

is used as a model plant due to its early growth habit and relatively short lifecycle, early asexual propagation from cuttings, callus or protoplasts, availability of large sets of mutants, amenity for cytogenetic analysis. Besides ornamental

also possesses medicinal value like it antioxidant activity and

MATERIALS AND METHODS

Washing and sterilization of explants vitro protocols, leaf explants

were collected from plants growing in Kashmir University Botanical Garden. In order to remove dirt and dust, the explants were thoroughly washed under running tap water followed by washing with detergent labolene and surfactant tween-20. The

removed by washing the explants with double distilled water. Then the leaf explants were treated with chemical sterilants2% sodium hypochlorite for 5 minutes under laminar air flow hood. This was followed by washing with autoclaved

Finally the leaf explants were inoculated on Murashige and Skoog (1962) medium.

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Medium and culture conditions Murashige and Skoog (1962) medium was gelled with 0.8% agar and supplemented with different concentrations of different carbon sources (sucrose, glucose, mannose, arabinose and maltose). The pH of the medium was adjusted to 5.8 prior to autoclaving at 121 ºC and 15 lb pressure. The cultures were finally incubated at 22±4 ºC and maintained under controlled growth conditions. RESULTS In the present study, the effect of different carbon sources (sucrose, glucose, mannose, arabinose and maltose) on callus production and subsequent shoot regeneration from leaf explant was investigated (Table 1). MS medium supplemented with sucrose as carbon source was found to be best where on maximum number of callus was regenerated with

subsequent maximum number of shoot regeneration within 11 days with 70 percent culture response (Fig.1 a). Callus and subsequent shooting was also observed on MS medium supplemented withglucose and maltose with 50 and 30 percent culture response within 18 and 24 days respectively (Fig. 1b, 1c). Keeping in view the results obtained from MS medium fortified with sucrose, the effect of different concentrations of sucrose i.e., 2%, 3% and 4% was seen on shoot regeneration (Table 2).MS medium supplemented with 3% sucrose was found to be best medium where on maximum number of shoots were regenerated in 70% cultures within 15 days (Fig.1d). Shoot regeneration was also obtained when 2 and 4 percent sucrose was added to MS medium with culture response of 30 and40% within 28 and 25 days respectively (Fig. 1e, 1f).

a) b) c)

d) e) f)

Fig.1: In vitro shoot regeneration from leaf explant on MS medium fortified with a) Sucrose b) Glucose c) Maltose

d) 3% sucrose e) 2% sucrose f) 4% sucrose

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Table 1: Effect of MS medium fortified with different carbon sources on shoot regeneration of Petunia hybrid:

Carbon

source

Callus

induction

No. of days

taken for

shooting

Sucrose + 11

Glucose + 18

Maltose + 24

mannose - -

Arabinose - -

- No response; + Positive response

Table 2: Effect of MS medium fortified with different concentrations of sucrose on shoot

regeneration of Petunia hybrid:

Carbon

source

Callus

inductio

n

No. Of days

taken for

shooting

2% Sucrose ++ 18

3% sucrose +++ 11

4% sucrose + 24

+ low amount; ++ moderate amount; +++ high amount

DISCUSSION The aim of present study was to investigate the effect of different carbon sources on the growth of Petunia hybrida. The present study reveals that sucrose at 3% concentration showed best results in terms of callus induction and shoot regeneration. Similarobtained by Marcelinaet al. in 2015. He observed the highest number of shoots in case of Prism Rose’, on MS medium supplemented with sucrose. Our results are also in accordance with that of Yistraet al., who also found in vitro

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Table 1: Effect of MS medium fortified with different carbon sources on shoot regeneration of

% Culture

response

70%

50%

30%

-

-

No response; + Positive response

Table 2: Effect of MS medium fortified with different concentrations of sucrose on shoot

regeneration of Petunia hybrid:

days

%

Culture

response

30%

70%

40%

+ low amount; ++ moderate amount; +++ high

The aim of present study was to investigate the effect of different carbon sources on the growth of Petunia

The present study reveals that sucrose at 3% concentration showed best results in terms of callus induction and shoot regeneration. Similar results were

. in 2015. He observed the highest number of shoots in case of Petunia cv.’ Prism Rose’, on MS medium supplemented with

Our results are also in accordance with that in vitropollen

germination of Petunia hybridacontaining sucrose as a carbon source(2014) also studied the impact of different carbohydrates and their concentrations on regeneration of Solanumviarumthat the maximum number of shoots and shoot length was obtained on MS medium supplemented with 4% fructose. Our results are in contrast with that of Raquin(1983)who observed maltose as a carbon source best for in vitro anther culture of petunia. REFERENCES 1. Ando, T., Ishikawa, N., Watanabe, H., Kokubun,

H., Yanagisawa, Y., Hashimoto, G., Marches, E and Suarez, E. A. (2005). Morphological study of Petunia integrifoliacomplex (Solnaceae). of Botany. 96: 887-900

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3. Krupa-Małkiewicz, M., Fornal, N., (2018). Effect of Carbohydrate source on propagation and rooting of two cultivars ofPetunia (Petunia × Atkinsiana D. Don). Aliment PiscZootech. 322:

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Petunia hybridaon MS medium containing sucrose as a carbon source. Mahadevet al. (2014) also studied the impact of different carbohydrates and their concentrations on in vitro

Solanumviarum (Dunal). He observed that the maximum number of shoots and shoot length was obtained on MS medium supplemented with 4% fructose. Our results are in contrast with that of Raquin(1983)who observed maltose as a carbon

anther culture of petunia.

Ando, T., Ishikawa, N., Watanabe, H., Kokubun, H., Yanagisawa, Y., Hashimoto, G., Marches, E and Suarez, E. A. (2005). Morphological study of

complex (Solnaceae). Annuals

Panathula, S.C., Naidu, V.C. Impact of differentcarbohydrates and

in vitro regeneration of —Animportant anticancer

American Journal of Plant

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(Petunia × Atkinsiana D. Don). Agric . 322:67-78.

Raquin, C., (1893). Utilization of different sugars in vitro anther culture of

Petunia. Zeitschrift Fur Pflanzenphysiologie.

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Garrido, D., Jacqueline Busscher, Tunen, J.A., (1998). Hexose Transport in

Growing Petunia Pollen Tubes and Characterization Specific, Putative Monosaccharide

DOI: https://doi.org/10.1104/.118-